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SUMMARY: Overexpression of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in various tumor tissues and cell lines was found to promote tumor cell proliferation, migration, and invasion. However, the role of MALAT1 in gastric cancer (GC) is still unclear. We aimed to investigate the correlation between long-chain non-coding RNAs (lncRNAs), MALAT1, MicroRNAs (miRNA) and vascular endothelial growth factor A (VEGFA) in gastric cancer and to disclose underlying mechanism. The correlation between MALAT1 levels and clinical features was analyzed by bioinformatics data and human samples. The expression of MALAT1 was down regulated in AGS cells to detect the cell proliferation, migration, and invasion characteristics, as well as the effects on signal pathways. Furthermore, we validated the role of MALAT1/miR-330-3p axis in GC by dual luciferase reporter gene assays. Expression of MALAT1 was higher in cancer tissues than in para-cancerous tissues. The high MALAT1 level predicted malignancy and worse prognosis. Down-regulation of MALAT1 expression in AGS cells inhibited cell proliferation, migration, and invasion by targeting VEGFA. By dual luciferase reporter gene assay and miR-330-3p inhibitor treatment, we demonstrate that MALAT1 sponged miR-330-3p in GC, leading to VEGFA upregulation and activation of the mTOR signaling pathway. The MALAT1/miR-330-3p axis regulates VEGFA through the mTOR signaling pathway and promotes the growth and metastasis of gastric cancer.
Se descubrió que la sobreexpresión del transcrito 1 de adenocarcinoma de pulmón asociado a metástasis (MALAT1) en varios tejidos tumorales y líneas celulares promueve la proliferación, migración e invasión de células tumorales. Sin embargo, el papel de MALAT1 en el cáncer gástrico (CG) aún no está claro. Nuestro objetivo fue investigar la correlación entre los ARN no codificantes de cadena larga (lncRNA), MALAT1, los microARN (miARN) y el factor de crecimiento endotelial vascular A (VEGFA) en el cáncer gástrico y revelar el mecanismo subyacente. La correlación entre los niveles de MALAT1 y las características clínicas se analizó mediante datos bioinformáticos y muestras humanas. La expresión de MALAT1 se reguló negativamente en las células AGS para detectar las características de proliferación, migración e invasión celular, así como los efectos sobre las vías de señales. Además, validamos el papel del eje MALAT1/miR- 330-3p en GC mediante ensayos de genes indicadores de luciferasa dual. La expresión de MALAT1 fue mayor en tejidos cancerosos que en tejidos paracancerosos. El alto nivel de MALAT1 predijo malignidad y peor pronóstico. La regulación negativa de la expresión de MALAT1 en células AGS inhibió la proliferación, migración e invasión celular al apuntar a VEGFA. Mediante un ensayo de gen indicador de luciferasa dual y un tratamiento con inhibidor de miR-330-3p, demostramos que MALAT1 esponjaba miR-330-3p en GC, lo que lleva a la regulación positiva de VEGFA y la activación de la vía de señalización mTOR. El eje MALAT1/miR-330-3p regula VEGFA a través de la vía de señalización mTOR y promueve el crecimiento y la metástasis del cáncer gástrico.
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Humains , Tumeurs de l'estomac/génétique , Tumeurs de l'estomac/anatomopathologie , Facteur de croissance endothéliale vasculaire de type A , Sérine-thréonine kinases TOR , ARN long non codant , ARN/génétique , Transduction du signal , Régulation de l'expression des gènes tumoraux , Mouvement cellulaire , Technique de Western , Apoptose , Gènes rapporteurs , Prolifération cellulaire , Réaction de polymérisation en chaine en temps réel , Invasion tumoraleRÉSUMÉ
Aim To analyze the differences in plasma biomarkers and metabolic pathways between Atractylodes chinensis and Atractylodes coreana after intervention in spleen deficiency rats, and discuss the spleen strengthening mechanism of the two from a non targeted metabolomics perspective. Methods A spleen deficiency model was established in SD rats using a composite factor method of improper diet, excessive fatigue, and bitter cold diarrhea. To determine the content of gastrointestinal and immunological indicators, UHPLC-QE-MS technology was used, combined with principal component analysis (PC A) and orthogonal projections to latent structures-discriminant analysis (OPLS-DA) methods to search for biomarkers in plasma of spleen deficiency rats, and metabolic pathways were induced using the Pathway database. Results After administration of Atractylodes chinensis and Atractylodes coreana, various indicators in plasma of spleen deficiency rats showed varying degrees of regression. Metabolomics analysis showed that Atractylodes chinensis and Atractylodes coreana respectively recalled 70 and 82 plasma differential metabolites. Atractylodes chinensis mainly regulated two metabolic pathways : "Glycine, serine, and threonine metabolism, and "Thiamine metabolism". Atractylodes coreana mainly regulated five metabolic pathways, "Glycine, serine, and threonine metabolism", "Thiamine metabolism, "Pyrimidine metabolism", "Butanoate metabolism", and "Riboflavin metabolism". Conclusions Both Atractylodes chinensis and Atractylodes coreana have certain regulatory effects on spleen deficiency rats, and their mechanism of action may be related to regulating metabolic pathways such as "Glycine, serine, and threonine metabolism, and "Thiamine metabolism"in spleen deficiency.
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OBJECTIVE@#To analyze the reliability of the Water Tank Scale for assessing recovery of motor function after spinal cord injury (SCI) in rats.@*METHODS@#Thirty-six adult female SD rats were randomly divided into SCI and sham-operated groups (n= 18). The recovery of the hind limb motor function was assessed using Water Tank scoring, BBB scoring, and motor-evoked potentials (MEP) at 1, 3, 5, 7, 14 and 21 days after SCI. MEP was used as the gold standard for analyzing and comparing differences between the two scoring methods.@*RESULTS@#The Water Tank scores of the rats were significantly higher than the BBB scores on day 3 (0.22±0.43 vs 0, P < 0.05) and also on days 5, 7 and 14 after SCI (0.67±0.49 vs 0.11±0.32, 4.33±1.19 vs 2.83±1.04, 8.61± 1.20 vs 7.06±1.0, P < 0.01). On day 21 after SCI, the scores of the Water Tank Scale of the rats did not significantly differ from the BBB scores (14.78±1.06 vs 14.50±1.47, P>0.05). Neurophysiological monitoring showed that both the Water Tank score and BBB score were significantly correlated with MEP latency, but the Water Tank score had a greater correlation coefficient with MEP latency (r=-0.90).@*CONCLUSION@#Compared with the BBB scale, Water Tank scoring allows more objective and accurate assessment of functional recovery of the spinal cord in early stages following SCI in rats, and can thus be used as a reliable method for assessing functional recovery of the hind limbs in rat models of acute SCI.
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Femelle , Animaux , Rats , Rat Sprague-Dawley , Reproductibilité des résultats , Traumatismes de la moelle épinière , EauRÉSUMÉ
This study aimed to parallelly investigate the cardioprotective activity of Cinnamomi Ramulus formula granules(CRFG) and Cinnamomi Cortex formula granules(CCFG) against acute myocardial ischemia/reperfusion injury(MI/RI) and the underlying mechanism based on the efficacy of "warming and coordinating the heart Yang". Ninety male SD rats were randomly divided into a sham group, a model group, CRFG low and high-dose(0.5 and 1.0 g·kg~(-1)) groups, and CCFG low and high-dose(0.5 and 1.0 g·kg~(-1)) groups, with 15 rats in each group. The sham group and the model group were given equal volumes of normal saline by gavage. Before modeling, the drug was given by gavage once a day for 7 consecutive days. One hour after the last administration, the MI/RI rat model was established by ligating the left anterior descending artery(LAD) for 30 min ischemia followed by 2 h reperfusion except the sham group. The sham group underwent the same procedures without LAD ligation. Heart function, cardiac infarct size, cardiac patho-logy, cardiomyocyte apoptosis, cardiac injury enzymes, and inflammatory cytokines were determined to assess the protective effects of CRFG and CCFG against MI/RI. The gene expression levels of nucleotide-binding oligomerization domain-like receptor family pyrin domain protein 3(NLRP3) inflammasome, apoptosis-associated speck-like protein containing a CARD(ASC), cysteinyl aspartate specific proteinase-1(caspase-1), Gasdermin-D(GSDMD), interleukin-1β(IL-1β), and interleukin-18(IL-18) were determined by real-time quantitative polymerase chain reaction(RT-PCR). The protein expression levels of NLRP3, caspase-1, GSDMD, and N-GSDMD were determined by Western blot. The results showed that both CRFG and CCFG pretreatments significantly improved cardiac function, decreased the cardiac infarct size, inhibited cardiomyocyte apoptosis, and reduced the content of lactic dehydrogenase(LDH), creatine kinase MB isoenzyme(CK-MB), aspartate transaminase(AST), and cardiac troponin Ⅰ(cTnⅠ). In addition, CRFG and CCFG pretreatments significantly decreased the levels of IL-1β, IL-6, and tumor necrosis factor-α(TNF-α) in serum. RT-PCR results showed that CRFG and CCFG pretreatment down-regulated the mRNA expression levels of NLRP3, caspase-1, ASC, and downstream pyroptosis-related effector substances including GSDMD, IL-18, and IL-1β in cardiac tissues. Western blot revealed that CRFG and CCFG pretreatments significantly decreased the protein expression levels of NLRP3, caspase-1, GSDMD, and N-GSDMD in cardiac tissues. In conclusion, CRFG and CCFG pretreatments have obvious cardioprotective effects on MI/RI in rats, and the under-lying mechanism may be related to the inhibition of NLRP3/caspase-1/GSDMD signaling pathway to reduce the cardiac inflammatory response.
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Mâle , Animaux , Rats , Rat Sprague-Dawley , Interleukine-18 , Lésion de reperfusion myocardique , Protéine-3 de la famille des NLR contenant un domaine pyrine , Facteur de nécrose tumorale alpha , Infarctus du myocarde , Caspase-1RÉSUMÉ
This study explored the effects of propofol on the activity of glutamatergic neurons in the paraventricular thalamus (PVT) and the underlying mechanisms at the molecular level using whole-cell patch-clamp techniques. Acute brain slices containing the PVT were obtained from 8 weeks old C57BL/6J mice. The electrophysiological characteristics of PVT neurons were recorded in current-clamp mode, then single-cell sequencing was used to identify neuronal types. The firing frequencies before, during, and after propofol or intralipid application were recorded as FB, FD and FW; and the membrane potentials were recorded as MPB and MPD. Picrotoxin (PTX) was used to block inhibitory gamma-aminobutyric acid type A (GABAA) receptors during the application of propofol at 10 μmol·L-1. Then, GABAA receptor-mediated spontaneous and miniature inhibitory postsynaptic currents (sIPSCs and mIPSCs) were recorded, and the effects of 10 μmol·L-1 propofol were investigated. The animal experiments were approved by the Medical Animal Administrative Committee of Shanghai Medical College Fudan University. The results showed that there were no significant differences in FB, FD and FW during intralipid and 2 μmol·L-1 propofol application. With propofol at 5, 10 and 20 μmol·L-1, FD decreased significantly when compared with FB, and FW increased significantly as compared with FD (P < 0.01). The inhibition degree of the three concentration groups was significantly different (P < 0.01). In addition, with propofol at 20 μmol·L-1, MPD hyperpolarized significantly (P < 0.01). In the presence of PTX, 10 μmol·L-1 propofol could not suppress the firing frequency of PVT glutamatergic neurons. Propofol at 10 μmol·L-1 prolonged the decay time of sIPSCs (P < 0.01) and mIPSCs (P < 0.05), and increased the amplitude (P < 0.01) of mIPSCs of PVT glutamatergic neurons. Together, these results indicate that propofol can inhibit the activity of PVT glutamatergic neurons in a concentration-dependent and reversible manner, and the effect is likely to be mediated by postsynaptic GABAA receptors.
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This data article presents data from the China Migrants Dynamic Survey (CMDS), a multi-wave, large-scale national cross-sectional survey of China's internal migrants from 2009 to 2018. The CMDS is an annual questionnaire survey conducted by the former National Health and Family Planning Commission (NHFPC) of the People's Republic of China. The respondents included in this survey are internal migrants over 15 years old. The sample was drawn from the China Migrant Population Information System, using multi-stage stratified sampling method and the probability proportional-to-size (PPS) cluster sampling strategy. Between 2009 and 2018, there were 1,527,650 internal migrants from 23 provinces, 5 autonomous regions and 4 municipalities participated in the surveys. The survey tools were a series of self-designed questionnaires with high inheritance and consistency designed and implemented by the NHFPC. The questionnaires mainly contain basic information of the respondents and their family members, migration status, healthcare or health behaviors, public health service utilization, social insurance, social integration, and family planning. The dataset is currently the most widely used survey data on China's internal migrants, offering information on migration patterns, healthcare and health behaviors, use of public health services, access to social security, social integration, and family planning, which are valuable for health planning, health decision-making, and health equity research.
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Humains , Adolescent , États-Unis , Services de planification familiale , Population de passage et migrants , Études transversales , Chine/épidémiologie , Enquêtes et questionnairesRÉSUMÉ
Objective To investigate the effect of glucocorticoids on the prognosis and inflammatory markers of patients with acute severe pancreatitis(SAP).Methods A retrospective collection was performed for SAP patients diagnosed and treated in the Department of General Hepatobiliary Surgery,the Second Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine from January 2020 to January 2023.SAP patients were divided into GC group and non-GC(NGC)group according to whether they used glucocorticoids(GC)or not.The two groups were compared with general data,inflammatory markers[neutrophil-lymphocyte ratio(NLR),monocyte-lymphocyte ratio(MLR),platelet-lymphocyte ratio(PLR),systemic inflammatory index(SII)and C-reactive protein-albumin ratio(CAR)],clinical events[continuous renal replacement therapy(CRRT),vasoactive drugs,ICU stay,total length of stay and survival)],and complications(organ failure,electrolyte imbalances,gastrointestinal bleeding and infection).Kaplan-Meier curve analysis of the relationship between GC and death in SAP patients.Logistic regression was used to explore the relationship between GC and death and organ failure.Results A total of 84 patients with SAP,40 in the GC group and 44 in the NGC group were included in the study.After treatment,NLR,MLR,PLR,SII and CAR in the GC group were significantly lower than those in the NGC group(P<0.05).There were no statistically significant differences between the GC and NGC groups in CRRT treatment,vasoactive drug use,surgery,ICU stay,total length of stay,electrolyte imbalance,gastrointestinal bleeding,pancreatic pseudocyst,abscess formation,pancreatic encephalopathy,and infectious complications(P>0.05);The mortality rate and organ failure incidence of patients in the GC group were significantly lower than those in the NGC group(P<0.05).The results of multivariate regression showed that the use of GC was an independent protective factor for death[OR=0.243,95% CI(0.062,0.948),P=0.042]and organ failure[OR=0.401,95% CI(0.165,0.976),P=0.044]in SAP patients.Conclusion GC can reduce the mortality rate and organ failure rate of SAP patients without increasing the incidence of additional complications,and has a high safety profile.
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Objective: To analyze the time distribution of the first positive nucleic acid detection in imported cases infected with SARS-CoV-2 reported nationwide in China and provide references for further improvement of the prevention and control of COVID-19 in international travelers. Methods: The data of imported cases infected with SARS-CoV-2 reported by provinces from 24 July 2020 and 23 July 2021 were collected for the analysis on the time distribution of the first positive nucleic acid detection after entering China. Results: A total of 7 199 imported cases infected with SARS-CoV-2 were reported in 28 provinces during 24 July 2020 to 23 July 2021. The median interval (Q1, Q3) from the entry to the first positive nucleic acid detection of SARS-CoV-2 was 1 (0, 5) day. The imported cases who had the first positive nucleic acid detections within 14 days and 14 days later after the entry accounted for 95.15% (6 850/7 199) and 4.85% (349/7 199) respectively. Among these cases, 3.65% (263/7 199), 0.88% (63/7 199) and 0.32% (23/7 199) had the first positive nucleic acid detections within 15-21 days, 22-28 days and 28 days later after the entry respectively. The proportion of asymptomatic infections were 47.24% (3 236/6 850) and 63.61% (222/349) among the cases who had the first positive nucleic acid detections within 14 days and 14 days later after the entry respectively. A total of 39.54% (138/349) of cases infected with SARS-CoV-2 with the first positive nucleic acid detections 14 days later after the entry had inter-provincial travel after the discharge of entry point isolation. Conclusions: About 5% of the imported cases infected with SARS-CoV-2 were first positive 14 days later after the entry. In order to effectively reduce the risk of domestic COVID-19 secondary outbreaks caused by imported cases, it is suggested to add a nucleic acid test on 8th -13th day after the entry.
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Humains , Infections asymptomatiques , COVID-19 , Chine/épidémiologie , Acides nucléiques , SARS-CoV-2RÉSUMÉ
italic>Gentiana crassicaulis Duthie ex Burk. is one of the plant sources of Gentianae Macrophyllae Radix (QinJiao). Gentiana tibetica King ex Hook. f. and Gentiana robusta King ex Hook. f. are relative species of G. crassicaulis. Due to the large intraspecific morphological variation, G. crassicaulis showed high morphological similarity with G. tibetica and G. robusta. And the distribution area of the three species overlaps to some extent, which makes it difficult to identify them. On the basis of morphological identification, the method of molecular identification of the three species was constructed in this study based on chloroplast genomes. The chloroplast genome of Gentiana tibetica is 148 765bp long, with LSC, SSC and IR 81 163 bp, 17 070 bp and 25 266 bp, respectively. The structure of the three is consistent. The chloroplast genome sequences of G. tibetica and G. crassicaulis are highly similar, and the number of variable sites is 9 (149 267 bp in total). Diagnostic SNP that could effectively identify the three species was screened and verified, and a dual-peak SNP detection method was established for the effective identification of each species and mixed samples. Our study provides basic data for the molecular identification of G. crassicaulis and its related species, and the arrangement of related Tibetan medicine.
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ObjectiveTo explore the effects of different treatment methods of "soothing liver, invigorating spleen, soothing liver and invigorating spleen, soothing liver first and then soothing liver and invigorating spleen, as well as invigorating spleen first and then soothing liver and invigorating spleen" on liver depression combined with liver injury in rats and their action mechanisms. MethodA six-week rat model of liver depression combined with liver injury was established by restraint stress and subcutaneous injection of carbon tetrachloride (CCl4, 5.89 g·kg-1, once every three days). At the same time, the drugs were given by gavage. Forty-eight male SD rats of clean grade were randomly divided into eight groups, namely the normal group, model group, bicyclol (0.2 g·kg-1) group, Sinisan (4.32 g·kg-1) group, Liu Junzitang (9.26 g·kg-1) group, Chaishao Liu Junzitang A (Chai A, soothing liver and invigorating spleen,13.57 g·kg-1) group, Chaishao Liu Junzitang B (Chai B, soothing liver first and then soothing liver and invigorating spleen, 13.57 g·kg-1) group, and Chaishao Liu Junzitang C (Chai C, invigorating spleen first and then soothing liver and invigorating spleen, 13.57 g·kg-1) group, with six rats in each group. The pathological changes in liver and colon tissues of each group were observed under light microscope and electron microscope. The serum biochemical indexes of the liver were detected using an automatic biochemical analyzer. The relative mRNA expression levels of Takeda G protein-coupled receptor 5 (TGR5) and intestinal mucosal zona occluden-1 (ZO-1), Occludin, and Claudin-1 in the liver and colon were detected by reverse-transcription polymerase chain reaction (RT-PCR). The positive expression rate of proliferating cell nuclear antigen (PCNA) in the colon was detected by immunohistochemistry. ResultCompared with normal group, the model group exhibited significantly elevated serum alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), and direct bilirubin (DBIL) (P<0.01), lowered TGR5 mRNA expression in liver tissue, up-regulated TGR5 mRNA expression in the colon tissue (P<0.05,P<0.01), and down-regulated ZO-1, Occludin, and tight junction protein-1 (Claudin-1) mRNA expression and PCNA in the colon tissue (P<0.01). Compared with the model group, bicyclol and Chai C remarkably decreased the levels of serum ALP, ALT, AST, TBIL, and DBIL (P<0.05,P<0.01), while Liu Junzitang, Chai A, Chai B, and Chai C significantly up-regulated the TGR5 mRNA expression in the liver and down-regulated its expression in the colon (P<0.01). Bicyclol, Chai A, Chai B, and Chai C enhanced the ZO-1 and Claudin-1 mRNA expression in the colon (P<0.05,P<0.01). Bicyclol, Sinisan, and Chai C increased PCNA expression (P<0.01). The comparison with the Chai C group showed that the TGR5 mRNA expression in the liver and ZO-1 mRNA expression in the colon of the bicyclol and Sinisan groups were lower, whereas the TGR5 mRNA expression in the colon was higher (P<0.01). However, the PCNA expression in the colon of the Liu Junzitang and Chai B groups declined significantly (P<0.05). ConclusionIn the presence of liver injury, invigorating spleen first helps to relieve the liver injury, and the efficacy of "spleen-invigorating" therapy in increasing the intestinal mucosal tight junction proteins and improving the gastrointestinal function is related to its activation of TGR5 to improve the intestinal mucosal barrier function, promote the renewal of intestinal stem cells, and drive the regeneration after injury.
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ObjectiveTo establish a traditional Chinese medicine (TCM) syndrome model with yin deficiency and internal heat, discuss the rationality of model evaluation, and analyze differentially expressed genes in multiple dimensions to explore the molecular mechanism-signaling pathways as well as key targets of Baihe Dihuangtang (BHDH) in treating depression with Yin deficiency and internal heat. MethodForty male SD rats were randomly divided into a blank control group,a model group,a fluoxetine group (positive drug),a BHDH group, and a Zhibai Dihuangtang group (positive drug for Yin deficiency and internal heat). The depression model with Yin deficiency and internal heat was induced by chronic unpredictable mild stress (CUMS)combined with Chinese herbal drugs with warm and heat nature. The model established was comprehensively evaluated by the detection of the basic condition, behavioral performance, and biochemical indicators of rats in each group. The differentially expressed genes were screened out by mRNA sequencing and underwent Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses. The protein-protein interaction (PPI) network was plotted and key genes were analyzed to explore the underlying mechanism of BHDH in treating depression with Yin deficiency and internal heat. ResultThe comparison of basic conditions, behavioral assays, energy metabolism, endocrine hormones, cytokines, and neurotransmitters showed that the model was properly induced. BHDH could significantly improve depression with Yin deficiency and internal heat by regulating the pathways related to the nervous system, endocrine system, and inflammatory and immune system. The key genes of the PPI network were Fos, Epha8, Npy2r, Htr2c, and Nr4a1. ConclusionUnder the guidance of TCM theories of treatment based on syndrome differentiation and etiology and pathogenesis,this study established a depression model with yin deficiency and internal heat in animals and evaluation system in accordance with the symptoms and signs of emotional diseases, and further confirmed the scientificity of the modeling method and the underlying mechanism of BHDH in interfering with depression with Yin deficiency and internal heat based on the results of mRNA sequencing.
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The key factors for producing the best quality Chinese herbal medicines are high-quality germplasm, suitable cultivation area and the proper processing methods for herbal raw materials. Gentiana crassicaulis in Gentiana (Sect. Cruciata), Gentianaceae is one of the original plants of the Chinese herb Qinjiao (Gentianae Macrophyllae Radix), and its type specimen was collected in Lijiang, Yunnan. There is a long planting history of the herb in this area. In this study a sampling plot was designated in these traditional planting areas. G. crassicaulis was planted and herbal raw materials were harvested from the plot. The raw materials were prepared locally and at a pharmaceutical factory in Shanghai using processing methods such as "sweating" or "no sweating", "slicing" or "no slicing" (whole root), and "stoving" or "no stoving" (air drying). The quality of all processed samples was evaluated. In addition, molecular markers were determined for identifying cultivated and wild samples from Lijiang, Yunnan. The results are as follows: ① Samples from the sampling plot and the field are taxonomically identified as Gentiana crassicaulis. ② A total of 270 sequences of trnC-GCA-petN, atpB-rbcL, psbN, ndhB-rps7 and ycf1 were obtained, and three genotypes were determined from the cultivated samples; the type III was shared by both cultivated and wild plants. Based on the molecular markers, a DNA barcoding method to identify cultivated and wild samples of G. crassicaulis from Lijiang, Yunnan was established. ③ Total content of loganic acid and gentiopicroside in all samples was ≥ 2.5%, and above the Chinese Pharmacopoeia (2020) limit. ④ In HPLC fingerprinting, 9 common peaks were assigned and similarity between all samples was > 0.999; and ⑤ In a PCA score plot all slice samples were clustered, while whole root samples were scattered. Therefore, our studies could provide basic data for optimizing the processing method, producing best quality Gentianae Macrophyllae Radix, and evaluating the quality of different ecotype varieties and the multiple origin of herbal medicines.
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Aim To investigate the anti-tumor effect of celastrol(CEL)on colorectal cancer and the possible targets/mechanisms. Methods The cytotoxic activities of CEL were evaluated against A549, HCT-116, HepG2 by CCK-8 method. Western blotting was used to detect the expression level of STAT3 and its upstream and downstream proteins(JAK2, Survivin, MCL-1)in HCT-116 cells before and after CEL treatment Flow cytometry was applied to assess CEL's apoptosis and cell-cycle arrest effect in HCT-116 cells. SPR detection and molecular docking analysis were performed to further assess the binding ability between CEL and STAT3 protein. Lastly, human colorectal cancer organoid culture was constructed to verify the anti-tumor effect of CEL. Results CEL showed significant cytotoxicity to A549(IC50 = 2.37±0.02 μmol·L-1), HCT-116(IC50 = 1.40±0.21 μmol·L-1)and HepG2(IC50 = 2.52±0.02 μmol·L-1). Additionally, CEL could effectively decrease the level of p-STAT3 and the downstream gene expression of STAT3(Survivin and MCL-1)in a concentration-dependent manner; however, CEL did not affect the total level of STAT3 and upstream kinases JAK2. Moreover, CEL could induce apoptosis of HCT-116 cells concentration-dependently and arrest the cell cycle. According to the SPR analysis, CEL showed a strong binding affinity with the KD value(the equilibrium dissociation constant)of 60.38 μmol·L-1. Molecular docking analysis also suggested that CEL bound to the SH2 domain of STAT3. Lastly, CEL showed much better activity than the positive drug oxaliplatin(L-OHP)on all the colorectal cancer organoids. Conclusions CEL shows a significant anti-colorectal cancer effect, potentially caused by a direct target inhibiting STAT3, inducing apoptosis, and blocking the cell cycle.
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Aim To determine the antiviral and anti-inflammatory effects of the recommended prescription for COVID-2019-lung-spleen qi deficiency(4-1)against in vitro infection of SARS-CoV-2 and common coronaviruses.Methods The main chemical substances of 4-1 were analyzed by LC-MS.The toxicity and antiviral effects of of 4-1 were detected by MTT and by CPE assay, respectively.The viral loads in cell supernatant and the expression of inflammatory factors induced by viral infection were determined by qRT-PCR.Results The recommended prescription 4-1 contained 94 chemical compounds, including flavonoids, steroids, sesquiterpenoids, and so on.The range of selection indexes for SARS-CoV-2 and common coronaviruses was 8.44±0.4952.26±2.3.This prescription could inhibit the proliferation of SARS-CoV-2, the expression of ACE2 and S mRNA, and down regulate IL-1α and CCL-5/RANTES at 10, 5, and 2.5 g•L-1 doses.Further, at doses of 20, 10 and 5 g•L-1, it could inhibit the proliferation of three common coronaviruses and suppress the overexpression of IL-6, CXCL-8/IL-8, CXCL-10/IP-10, TNF-α, IFN-α, CCL-2/MCP-1, MIG and CCL-5/RANTES induced by OC43/229E infection.The inhibitory effects were dose-dependent.Conclusions The prescription 4-1 has antiviral and anti-inflammation effects against multiple coronaviruses.This study provides the research basis for the treatment of common respiratory viral infections and emerging infectious diseases such as COVID-19 by using traditional Chinese medicine.
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Objective:Powders and decocted powders account for about 1/3 in the Catalogue of Ancient Famous Classical Formulas (the First Batch), and have a very important position. Determination of preparation technology and particle size in the pulverization process is the key step in the research and development of powders and decocted powders following the original methods. However, there are many terms describing the preparation technology and particle size of powders and decocted powders in ancient Chinese medical books, and the parameters are not clear. Due to the lack of unified basis of particle size, the existing research results have not formed a uniform consensus. Based on ancient textual researches and experimental results, this article discusses the particle size of decocted powders and powders. Method:Through textual researches of the preparation technology and particle size of powders and decocted powders and powder classification in the 2020 edition of Chinese Pharmacopoeia, the specifications of pulverized particle size were suggested. In addition, Xiebaisan and Danggui Buxuetang were taken as examples to investigate the influence of different particle sizes (4, 10, 24 mesh) on the preparation process of decocted powders and the obtained decoction. Result:The particle size of 4 mesh was equivalent to that of ancient as big as hemp bean. The contents of index components in Xiebaisan and Danggui Buxuetang with particle size of 4 mesh were higher than that of 10 mesh and 24 mesh, but the particle size of 50 mesh was too fine to be filtered. Conclusion:The suggested particle sizes of powders and decocted powders are recommended as Cumo is the power through 10-mesh sieve, Mo is the power through 24-mesh sieve, Ximo is the power through 80-mesh sieve, as big as hemp bean is the power through 4-mesh sieve and not through 10-mesh sieve.
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Objective:To investigate the characteristics of skin diseases in patients with spinal cord injury (SCI). Methods:From January, 2012 to December, 2016, all the inpatients with SCI were collected through hospital information system, in which, the patients with skin problems referred to the dermatologists were screened. Their general demographic characteristics, the time of skin onset since SCI, segment of SCI, level of SCI, the distribution of skin lesion, and the dermatological diagnosis were all recorded. The number of skin diseases in each SCI patient was counted. Results:A total of 3152 inpatients with SCI were included, out of whom, 554 patients were referred to the dermatologists, and 785 person-times dermatological conditions were diagnosed. Among the 554 patients, the consultation rate of male patients was significantly higher than females (χ2 = 13.284, P < 0.001); the consultation rate of aged 18~35 years and aged 36~50 years groups was higher than aged less than 18 years and aged more than 35 years groups (χ2 = 15.994, P < 0.01); the appearence of the skin lesions within six months post-SCI was significantly higher than more than six months post-SCI (χ2 = 123.725, P < 0.001); the consultation rate of patients with cervical SCI was higher than those with thoracic and lumbosacral SCI (χ2 = 10.482, P < 0.01), and the consultation rate of tetraplegic patients was higher than paraplegia patients (χ2 = 9.172, P < 0.01). A total of 385 patients suffered one dermatological condition, while 169 patients suffered two or more dermatological conditions. Among them, men (χ2 = 6.108, P < 0.05), patients with cervical SCI (χ2 = 8.592, P < 0.05) and tetraplegic patients (χ2 = 8.592, P < 0.05) were more likely to suffer two or more dermatological conditions. Allergic (252 cases), infectious (186 cases) and sebaceous gland related (169 cases) skin diseases were the most common skin problems. Conclusion:Male, cervical SCI and tetraplegic patients are more likely to suffer two or more skin diseases. It is necessary to strengthen the prevention of skin diseases common in SCI patients.
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Objective:To explore the regulatory effect of Huadu Sanyinfang on phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt)/transcription factor nuclear factor-<italic>κ</italic>B (NF-<italic>κ</italic>B) in triple-negative breast cancer (TNBC) patients with qi-deficiency constitution based on the differential expression of miRNA. Method:Based on previous research results, this study conducted the bioinformatics analysis to predict the target genes responsible for regulating the differential expression of miRNA between patients with qi-deficiency constitution and those with moderate constitution, which were intersected with TNBC target genes. The resulting intersection targets were then subjected to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and protein-protein interaction (PPI) network analysis to obtain the key pathways and target genes for differentially expressed miRNA in regulating TNBC. TNBC patients with Qi-deficiency constitution were treated with Huadu Sanyinfang for three years after they completed the standard Western medical treatment. The peripheral blood of the patients was sampled before and after medication for detecting gene expression in the key pathways. Result:The comparison between patients with Qi-deficiency constitution and those with moderate constitution revealed 49 differentially expressed miRNAs (16 up-regulated and 33 down-regulated), which regulated 1 445 TNBC target genes. As demonstrated by PPI and KEGG pathway enrichment analysis, the key genes were mainly tumor protein p53 (TP53), Akt1, epidermal growth factor receptor (EGFR), mitogen-activated protein kinase 3 (MAPK3), vascular endothelial growth factor A (VEGRA), and tumor necrosis factor (TNF). The key pathways included PI3K/Akt, MAPK, and RAS signaling pathways. A total of 11 TNBC patients with qi-deficiency constitution were enrolled. Compared with the situations before treatment, the expression levels of p105 subunit of NF-κB (NF-<italic>κ</italic>B1) and Akt1 in the PI3K/Akt signaling pathway were down-regulated after medication, while the levels of catalytic subunit alpha of PI3K (PIK3CA) and B-cell lymphoma-xL (Bcl-xL) were up-regulated. The differences in NF-<italic>κ</italic>B1 and Akt1 expression were statistically significant. Conclusion:Huadu Sanyinfang is able to affect the gene expression of PI3K/Akt/NF-<italic>κ</italic>B signaling pathway in TNBC patients with Qi-deficiency constitution. Specifically, it down-regulates NF-<italic>κ</italic>B1 and Akt1 expression and up-regulates PIK3CA and Bcl-xL.
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Chaetocin is a natural metabolite product with various biological activities and pharmacological functions isolated from Chaetomium species fungi belonging to the thiodiketopyrazines. Numerous studies have demonstrated a wide range of antitumor activities of chaetocin in vitro and in vivo. Several studies have demonstrated that chaetocin sup?presses the growth and proliferation of various tumour cells by regulating multiple signalling pathways related to tumour initiation and progression, inducing cancer cell apoptosis (intrinsic and extrinsic), enhancing autophagy, inducing cell cycle arrest, as well as inhibiting tumour angiogenesis, invasion and migration. The antitumor effects and molecular mechanisms of chaetocin are reviewed and analysed in this paper, and the prospective applications of chaetocin in cancer prevention and therapy are also discussed. Our review provides the theoretical basis for exploiting the clinical applica?tion of chaetocin in cancer treatment.
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As two original plants of Tibetan herb Jieji, Gentiana waltonii Burk. and Gentiana lhassica Burk. belong to Section Cruciata of Gentiana, Gentianaceae. Here, we report on whole chloroplast genome sequences in the alpine species, respectively, and the features of plastomes were investigated. The plastome of G. waltonii is 148 705 bp long (148 652 bp in G. lhassica) and encodes 112 genes, including 78 protein-coding genes, 30 transfer RNA genes, and 4 ribosomal RNA genes. Two pseudogenes, namely ψrps16 and ψinfA, were found in plastomes. In addition, two novel loci were detected, and a species-specific polymerase chain reaction assay was developed for differentiating G. waltonii and G. lhassica from 10 alpine species in Section Cruciata. Gentiana. Our study provides basic data for identifying Tibetan herbs, alpine species conservation and molecular phylogenetic studies of Gentiana and Gentianaceae.
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italic>Gentiana crassicaulis Duthie ex Burk. in Gentiana (Sect. Cruciata), Gentianaceae, is one of the original plants of both Gentianae Macrophyllae Radix and Tibetan herb Jie-Ji Na-Bao, which contain such bioactive iridoids as gentiopicroside, loganic acid and others. In this study, based on previous work, the transcriptome of G. crassicaulis was sequenced and analyzed to construct transcriptome databases of roots, stems, leaves and flowers. qRT-PCR verification was conducted for parts of unigenes that may be key enzymes in the pathway of iridoid biosynthesis. The results are as follows: ① a total of 159 534 unigenes were obtained, with an average length of 679 bp. According to the functional classification of GO, unigenes can be divided into 3 categories with 67 branches. The unigenes were aligned in the KOG database and were classified into 25 categories according to function. ② In the KEGG database, 215 unigenes were implicated in 20 standard secondary metabolism pathways. The analysis shows that 305 unigenes encoded 28 key enzymes in the pathway of iridoid biosynthesis, and their expression in different organs is different; and ③ qRT-PCR was approximately consistent with RNA-Seq results. The 7 annotated unigenes identified in this study, HMGS, DXS, MCS, GPPS, G10H, 7-DLNGT and STR, all had higher relative expression levels in the above-ground parts (stem, leaf and flower) than in the underground part (root). Iridoids are common active and index components of such traditional Chinese medicines as Qinjiao, Longdan, Dangyao, and Qingyedan, among others. Therefore, this work provides basic scientific data for further development including obtaining active components or intermediates through biotechnology, exploring the accumulation of effective components, evaluating the quality of different ecotype varieties, and identifying authentic biosynthesis pathways of medicinal materials.